LOCUS       Exported                5434 bp ds-DNA    circular SYN 17-OCT-2014
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    piG0071_sensor_pluxRRR12y_bxb1_mCherry
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 5434)
  AUTHORS   Daniel Gerngross
  TITLE     Direct Submission
  JOURNAL   Exported 17 Oct 2014 from SnapGene 2.5.0
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5434
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     terminator      1..126
                     /note="T0"
     terminator      27..121
                     /note="lambda t0 terminator"
                     /note="transcription terminator from phage lambda"
     misc_feature    127..196
                     /note="R2oDNA spacer"
                     /note="R2oDNA 70bp spacer 13"
     terminator      203..230
                     /note="T7Te"
                     /note="phage T7 early transcription terminator"
     misc_feature    complement(238..308)
                     /note="taR12y"
     promoter        complement(309..363)
                     /note="lux rR (R0062)"
     misc_feature    364..433
                     /note="spacer (B0040)"
     promoter        434..488
                     /note="lux rR (R0062)"
     3'UTR           489..540
                     /note="crR12y"
     RBS             523..534
                     /note="RBS (B0034)"
     CDS             541..2043
                     /codon_start=1
                     /note="bxb1"
                     /translation="MRALVVIRLSRVTDATTSPERQLESCQQLCAQRGWDVVGVAEDLD
                     VSGAVDPFDRKRRPNLARWLAFEEQPFDVIVAYRVDRLTRSIRHLQQLVHWAEDHKKLV
                     VSATEAHFDTTTPFAAVVIALMGTVAQMELEAIKERNRSAAHFNIRAGKYRGSLPPWGY
                     LPTRVDGEWRLVPDPVQRERILEVYHRVVDNHEPLHLVAHDLNRRGVLSPKDYFAQLQG
                     REPQGREWSATALKRSMISEAMLGYATLNGKTVRDDDGAPLVRAEPILTREQLEALRAE
                     LVKTSRAKPAVSTPSLLLRVLFCAVCGEPAYKFAGGGRKHPRYRCRSMGFPKHCGNGTV
                     AMAEWDAFCEEQVLDLLGDAERLEKVWVAGSDSAVELAEVNAELVDLTSLIGSPAYRAG
                     SPQREALDARIAALAARQEELEGLEARPSGWEWRETGQRFGDWWREQDTAAKNTWLRSM
                     NVRLTFDVRGGLTRTIDFGDLQEYEQHLRLGSVVERLHTGMS"
     misc_feature    2047..2066
                     /note="35 nt upstream of ATG from pET vectors"
                     /note="35 nt upstream of ATG from pET vectors (pET RBS) -->
                     introduction of SphI site for library cloning (SphI-HF!!)"
     misc_feature    2082..2795
                     /note="mCherry"
     misc_feature    2097..2102
                     /note="introduction"
                     /note="introduction of two point mutations to disrupt 
                     second inframe RBS in the mCherry gene (2x GAG --> GAA for 
                     E)"
     terminator      2860..2976
                     /note="T1"
     terminator      2869..2955
                     /gene="Escherichia coli rrnB"
                     /note="T1"
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2983..3002
                     /note="AB179"
     primer_bind     3168..3187
                     /note="AB5"
     rep_origin      3321..4148
                     /direction=RIGHT
                     /note="p15A"
     rep_origin      3569..3571
                     /direction=RIGHT
                     /note="P15 ori from pACYC184"
     rep_origin      complement(3572..4116)
                     /direction=LEFT
                     /note="p15A ori"
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     primer_bind     4220..4239
                     /note="AB180"
     oriT            4250..4358
                     /note="incP origin of transfer"
     CDS             complement(4514..5329)
                     /note="KamR"
ORIGIN
        1 aaatcgtaat tattggggac ccctggattc tcaccaataa aaaacgcccg gcggcaaccg
       61 agcgttctga acaaatccag atggagttct gaggtcatta ctggatctat caacaggagt
      121 ccaagatatt cgtcacagta gttcaacctc agatacagaa accgtagatt gggattcagg
      181 aagttaccct attgtttata aacgcagaaa ggcccacccg aaggtgagcc agtgtgatct
      241 agagatatat ggtagtagta agttaatttt cattaaccac cactaccaat cacctcctgg
      301 atttagcttt tattcgacta taacaaacca ttttcttgcg taaacctgta cgatcctaca
      361 ggtaggttct gttaagtaac tgaacccaat gtcgttagtg acgcttacct cttaagaggt
      421 cactgaccta acaacctgta ggatcgtaca ggtttacgca agaaaatggt ttgttatagt
      481 cgaataaaga attctaccat tcacctcttg gatttagcta ttaaagagga gaaaggtacc
      541 gtgagagccc tggtagtcat ccgcctgtcc cgcgtcaccg atgctacgac ttcaccggag
      601 cgtcagctgg agtcttgcca gcagctctgc gcccagcgcg gctgggacgt cgtcggggta
      661 gcggaggatc tggacgtctc cggggcggtc gatccgttcg accggaagcg cagaccgaac
      721 ctggcccggt ggctagcgtt cgaggagcaa ccgtttgacg tgatcgtggc gtaccgggta
      781 gatcggttga cccgatcgat ccggcatctt cagcagctgg tccactgggc cgaggaccac
      841 aagaagctgg tcgtctccgc gaccgaagcg cacttcgata cgacgacgcc gtttgcggcg
      901 gtcgtcatcg cgcttatggg aacggtggcg cagatggaat tagaagcgat caaagagcgg
      961 aaccgttcgg ctgcgcattt caatatccgc gccgggaaat accgaggatc cctgccgccg
     1021 tggggatacc tgcctacgcg cgtggacggg gagtggcggc tggtgccgga ccctgtgcag
     1081 cgagagcgca tcctcgaggt gtatcaccgc gtcgtcgaca accacgagcc gctgcatctg
     1141 gtggcccacg acctgaaccg gcgtggtgtc ctgtcgccga aggactactt cgcgcagctg
     1201 caaggccgcg agccgcaggg ccgggagtgg tcggctaccg cgctgaagcg atcgatgatc
     1261 tccgaggcga tgctcgggta cgcgactctg aacggtaaga ccgtccgaga cgacgacgga
     1321 gccccgctgg tgcgggctga gccgatcctg acccgtgagc agctggaggc gctgcgcgcc
     1381 gagctcgtga agacctcccg ggcgaagccc gcggtgtcta ccccgtcgct gctgctgcgg
     1441 gtgttgttct gcgcggtgtg cggggagccc gcgtacaagt tcgccggggg aggacgtaag
     1501 cacccgcgct accgctgccg ctcgatgggg ttcccgaagc actgcgggaa cggcacggtg
     1561 gcgatggccg agtgggacgc gttctgcgag gagcaggtac tggatctgct cggggacgcg
     1621 gagcgtctgg agaaagtctg ggtagcgggc tcggactccg cggtcgaact cgcggaggtg
     1681 aacgcggagc tggtggacct gacgtcgctg atcggctccc cggcctaccg ggcgggctct
     1741 ccgcagcgag aagcactgga tgcccgtatt gcggcgctgg ccgcgcggca agaggagctg
     1801 gagggcctgg aggctcgccc gtctggctgg gagtggcgcg agaccgggca gcggttcggg
     1861 gactggtggc gggagcagga caccgcggca aagaacacct ggcttcggtc gatgaacgtt
     1921 cggctgacgt tcgacgtccg cggcgggctg actcgcacga tcgacttcgg ggatcttcag
     1981 gagtacgagc agcatctcag gctcggcagc gtggtcgaac ggctacacac cgggatgtcg
     2041 taataaataa ttttgtttaa ctttaatgag gaggtatacg catgcatagc aagggcgaag
     2101 aagataacat ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg gagggctccg
     2161 tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc
     2221 agaccgccaa gctgaaggtg accaagggtg gccccctgcc cttcgcctgg gacatcctgt
     2281 cccctcagtt catgtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact
     2341 acttgaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg
     2401 gcggcgtggt gaccgtgacc caggactcct ccttgcagga cggcgagttc atctacaagg
     2461 tgaagctgcg cggcaccaac ttcccctccg acggccccgt aatgcagaag aagaccatgg
     2521 gctgggaggc ctcctccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca
     2581 agcagaggct gaagctgaag gacggcggcc actacgacgc tgaggtcaag accacctaca
     2641 aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt caacatcaag ttggacatca
     2701 cctcccacaa cgaggactac accatcgtgg aacagtacga acgcgccgag ggccgccact
     2761 ccaccggcgg catggacgag ctgtacaaga ggccttaata agaattcgcg cggccgcggc
     2821 ctaggcggcc tcctgtgtga aattgttatc cgctttaatt aaaggcatca aataaaacga
     2881 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
     2941 ctgagtagga caaatccgcc gccctagaca gctgggcgcg ccaaatacgc ccggtagtga
     3001 tcttatttca ttatggtgaa agttggaacc tcttacgtgc cgatcaacgt ctcattttcg
     3061 ccaaaagttg gcccagggct tcccggtatc aacagggaca ccaggattta tttattctgc
     3121 gaagtgatct tccgtcacag gtatttattc ggcgcaaagt gcgtcgggtg atgctgccaa
     3181 cttactgatt tagtgtatga tggtgttttt gaggtgctcc agtggcttct gtttctatca
     3241 gctgtccctc ctgttcagct actgacgggg tggtgcgtaa cggcaaaagc accgccggac
     3301 atcagcgcta gcggagtgta tactggctta ctatgttggc actgatgagg gtgtcagtga
     3361 agtgcttcat gtggcaggag aaaaaaggct gcaccggtgc gtcagcagaa tatgtgatac
     3421 aggatatatt ccgcttcctc gctcactgac tcgctacgct cggtcgttcg actgcggcga
     3481 gcggaaatgg cttacgaacg gggcggagat ttcctggaag atgccaggaa gatacttaac
     3541 agggaagtga gagggccgcg gcaaagccgt ttttccatag gctccgcccc cctgacaagc
     3601 atcacgaaat ctgacgctca aatcagtggt ggcgaaaccc gacaggacta taaagatacc
     3661 aggcgtttcc cctggcggct ccctcgtgcg ctctcctgtt cctgcctttc ggtttaccgg
     3721 tgtcattccg ctgttatggc cgcgtttgtc tcattccacg cctgacactc agttccgggt
     3781 aggcagttcg ctccaagctg gactgtatgc acgaaccccc cgttcagtcc gaccgctgcg
     3841 ccttatccgg taactatcgt cttgagtcca acccggaaag acatgcaaaa gcaccactgg
     3901 cagcagccac tggtaattga tttagaggag ttagtcttga agtcatgcgc cggttaaggc
     3961 taaactgaaa ggacaagttt tggtgactgc gctcctccaa gccagttacc tcggttcaaa
     4021 gagttggtag ctcagagaac cttcgaaaaa ccgccctgca aggcggtttt ttcgttttca
     4081 gagcaagaga ttacgcgcag accaaaacga tctcaagaag atcatcttat taatcagata
     4141 aaatatttct agcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     4201 ggttccgcgc acatttcccc gaaaagtgcc acctgctagg gccggcctac ggccagcctc
     4261 gcagagcagg attcccgttg agcaccgcca ggtgcgaata agggacagtg aagaaggaac
     4321 acccgctcgc gggtgggcct acttcaccta tcctgcccgg ctgacgccgt tggatacacc
     4381 aaggaaagtc tacacgaacc ctttggcaaa atcctgtata tcgtgcgaaa aaggatggat
     4441 ataccgaaaa aatcgctata atgaccccga agcagggtta tgcagcggaa aaggaccctt
     4501 gtccaattaa ttattagaaa aattcatcca gcatcagatg aaattgcagt ttgttcatat
     4561 ccggattatc aatgccatat ttctgaaaca gacgtttttg caggctcggg ctaaattcgc
     4621 ccaggcagtt ccacagaatg gccagatcct gataacgatc cgcaatgccc acacggccca
     4681 catcaatgca gccaatcagt ttgccttcat cgaaaatcag gttatccagg ctaaaatcgc
     4741 cgtgggtcac cacgctatcc gggctaaacg gcagcagttt atgcatttct ttccacacct
     4801 gttccaccgg ccagccgtta cgttcatcat caaaatcgct cgcatccacc aggccgttgt
     4861 tcatacggct ctgcgcctgg gccagacgaa acacacgatc gctgttaaac gggcagttgc
     4921 acaccggaat gctatgcaga cgacgcagaa acacggccag cgcatccaca atgttttcgc
     4981 cgctatccgg atattcttcc agcacctgaa acgcggtttt gcccggaatc gcggtggtca
     5041 gcagccacgc atcatccggg gtgcgaataa aatgtttaat ggtcggcagc ggcataaatt
     5101 cggtcagcca gttcagacgc accatttcat cggtcacatc gttcgccacg ctgcctttgc
     5161 catgtttcag aaacagttcc ggcgcatccg gtttgccata cagacgataa atggtcgcgc
     5221 cgctctgacc cacgttatca cgcgcccatt tatagccata cagatccgca tccatgttgc
     5281 tgttcagacg cggacggcta cagctcgttt cacgctgaat atggctcata acaccccttg
     5341 tattactgtt tatgtaagca gacagtttta ttgttcatga tgatatattt ttatcttgtg
     5401 caatgtaaca tcagagattt tgagacacaa attt
//
